Peptide-containing alpha-ketoamide cysteine and serine protease inhibitors

ABSTRACT

This invention relates to peptide-containing α-ketoamide inhibitors of cysteine and serine proteases, methods for making these compounds, and methods for using the same.

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit of U.S. Provisional ApplicationSerial No. 60/061,309, filed Oct. 7, 1997, the disclosure of which ishereby incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

[0002] This invention relates to peptide-containing α-ketoamideinhibitors of cysteine and serine proteases, methods for making thesecompounds, and methods for using the same.

BACKGROUND OF THE INVENTION

[0003] Numerous cysteine and serine proteases have been identified inhuman tissues. A “protease” is an enzyme which degrades proteins intosmaller components (peptides). The terms “cysteine protease” and “serineprotease” refer to proteases which are distinguished by the presencetherein of a cysteine or serine residue which plays a critical role inthe catalytic process. Mammalian systems, including humans, normallydegrade and process proteins via a variety of enzymes including cysteineand serine proteases. However, when present at elevated levels or whenabnormally activated, cysteine and serine proteases may be involved inpathophysiological processes.

[0004] For example, calcium-activated neutral proteases (“calpains”)comprise a family of intracellular cysteine proteases which areubiquitously expressed in mammalian tissues. Two major calpains havebeen identified; calpain I and calpain II. While calpain II is thepredominant form in many tissues, calpain I is thought to be thepredominant form in pathological conditions of nerve tissues. Thecalpain family of cysteine proteases has been implicated in manydiseases and disorders, including neurodegeneration, stroke,Alzheimer's, amyotrophy, motor neuron damage, acute central nervoussystem injury, muscular dystrophy, bone resorption, plateletaggregation, cataracts and inflammation. Calpain I has been implicatedin excitatory amino-acid induced neurotoxicity disorders includingischemia, hypoglycemia, Huntington's Disease, and epilepsy. Thelysosomal cysteine protease cathepsin B has been implicated in thefollowing disorders: arthritis, inflammation, myocardial infarction,tumor metastasis, and muscular dystrophy. Other lysosomal cysteineproteases include cathepsins C, H, L and S. Interleukin-1β convertingenzyme (“ICE”) is a cysteine protease which catalyzes the formation ofinterleukin-1β. Interleukin-1β is an immunoregulatory protein implicatedin the following disorders: inflammation, diabetes, septic shock,rheumatoid arthritis, and Alzheimer's disease. ICE has also been linkedto apoptotic cell death of neurons, which is implicated in a variety ofneurodegenerative disorders including Parkinson's disease, ischemia, andamyotrophic lateral sclerosis (ALS).

[0005] Cysteine proteases are also produced by various pathogens. Thecysteine protease clostripain is produced by Clostridium histolyticum.Other proteases are produced by Trypanosoma cruzi, malaria parasitesPlasmodium falciparum and P. vinckei and Streptococcus. Hepatitis Aviral protease HAV C3 is a cysteine protease essential for processing ofpicornavirus structural proteins and enzymes.

[0006] Exemplary serine proteases implicated in degenerative disordersinclude thrombin, human leukocyte elastase, pancreatic elastase, chymaseand cathepsin G. Specifically, thrombin is produced in the bloodcoagulation cascade, cleaves fibrinogen to form fibrin and activatesFactor VIII; thrombin is implicated in thrombophlebitis, thrombosis andasthma. Human leukocyte elastase is implicated in tissue degenerativedisorders such as rheumatoid arthritis, osteoarthritis, atherosclerosis,bronchitis, cystic fibrosis, and emphysema. Pancreatic elastase isimplicated in pancreatitis. Chymase, an enzyme important in angiotensinsynthesis, is implicated in hypertension, myocardial infarction, andcoronary heart disease. Cathepsin G is implicated in abnormal connectivetissue degradation, particularly in the lung.

[0007] Given the link between cysteine and serine proteases and variousdebilitating disorders, compounds which inhibit these proteases would beuseful and would provide an advance in both research and clinicalmedicine. The present invention is directed to these, as well as other,important ends.

SUMMARY OF THE INVENTION

[0008] The present invention is directed to selected peptide-containingα-ketoamide inhibitors of cysteine and serine proteases represented bythe general formula I:

[0009] wherein:

[0010] Q has the formula G-B—(CHR⁴)_(v) where R⁴ is independently H oralkyl having from 1 to 4 carbons;

[0011] v is 0, 1, or 2;

[0012] B is selected from the group consisting of C(═O), OC(═O),S(═O)_(m), CH₂, a bond, NR⁵C(═O), S(═O)_(m)-A-C(═O), and C(═O)-A-C(═O),where R⁵ is H or lower alkyl;

[0013] m is 0, 1, or 2;

[0014] A is lower alkylene or cycloalkylene, optionally substituted withone or more halogen atoms, aryl, or heteroaryl groups;

[0015] M is a carbon atom;

[0016] G is selected from the group consisting of H, a blocking group,lower alkyl, lower alkenyl, aryl having from about 6 to about 14carbons, heterocyclyl having from about 5 to about 14 ring atoms,heterocycloalkyl having from about 5 to about 14 ring atoms, arylalkylhaving from about 7 to about 15 carbons, heteroarylalkyl, andarylheteroalkyl wherein the aryl portion can be unfused or fused withthe heteroalkyl ring, said alkyl, aryl, heterocyclyl, heterocycloalkyl,arylalkyl, heteroarylalkyl, and arylheteroalkyl groups being optionallysubstituted with one or more J groups;

[0017] J is selected from the group consisting of halogen, CN, nitro,lower alkyl, cycloalkyl, heterocycloalkyl, heteroalkyl, halogenatedalkyl, aryloxyalkyl, alkylthio, alkylsulfonyl, aryl, heteroaryl,arylalkyl, arylalkyloxy, arylsulfonyl, heteroarylsulfonyl,alkoxycarbonyl, alkoxyalkyl, acyl, alkoxy, hydroxy, carboxy,hydroxyalkyl, amino, alkylamino, and aminoalkyl, said amino group orsaid amino group of said aminoalkyl or alkylamino group being optionallysubstituted with an acyl group, an alkoxy group, or with 1 to 3 aryl,lower alkyl, cycloalkyl, or alkoxyalkyl groups; and said aryl,heteroaryl, heterocycloalkyl, and heteroalkyl groups being furtheroptionally substituted by a J group;

[0018] each Aaa is independently an amino acid which optionally containsone or more blocking groups;

[0019] n is 0, 1, 2, or 3;

[0020] R¹ and R² are independently selected from the group consisting ofH, alkyl having from one to about 6 carbons, arylalkyl having from about7 to about 15 carbons, heteroalkyl in which the ring contains from about5 to about 14 ring atoms, heteroarylalkyl in which the heteroaryl ringcontains from about 5 to about 14 ring atoms, alkoxyalkyl, a side chainof a naturally occurring amino acid in the R or S configuration, and(CH₂)_(p)NH-L, said alkyl, arylalkyl, heteroalkyl, heteroarylalkyl, andalkoxyalkyl groups being optionally substituted with one or more Jgroups;

[0021] p is 0, 1, 2, or 3;

[0022] L is selected from the group consisting of alkoxycarbonyl havingfrom 2 to about 7 carbons, arylalkoxycarbonyl in which the arylalkoxygroup contains about 7 to about 15 carbons, and S(═O)₂R⁶;

[0023] R⁶ is selected from the group consisting of lower alkyl, and arylhaving from about 6 to about 14 carbons;

[0024] R³ is selected from the group consisting of H, alkyl having fromone to about 6 carbons, arylalkyl having from about 7 to about 15carbons, heteroalkyl in which the ring contains from about 5 to about 14ring atoms, heteroarylalkyl in which the heteroaryl ring contains fromabout 5 to about 14 ring atoms, alkoxyalkyl, a side chain of a naturallyoccurring amino acid in the R or S configuration, (CH₂)_(p)NH-L,C(═O)R⁷, S(═O)₂R⁷, a blocking group, and when combined with the carbonatom to which R¹ is attached an alkylene group having from 2 to 5carbons, said alkylene group being optionally substituted with a groupselected from the group consisting of aryl, azide, CN, a protected aminogroup, and OSO₂-aryl, said alkyl, arylalkyl, heteroalkyl,heteroarylalkyl, and alkoxyalkyl groups being optionally substitutedwith one or more J groups;

[0025] R⁷ is selected from the group consisting of aryl having fromabout 6 to about 14 carbons, heteroaryl having from about 5 to about 14ring atoms, arylalkyl having from about 7 to about 15 carbons, alkylhaving from 1 to about 10 carbons, said aryl, heteroaryl, arylalkyl andalkyl groups being optionally substituted with one or more J groups,heteroalkyl having from 2 to about 7 carbons, alkoxy having from about 1to about 10 carbons, and amino optionally substituted with 1 or morealkyl groups;

[0026] q is 0 or 1;

[0027] Z is selected from the group consisting of C(═O)C(═O)NH—X-A¹-Kand

[0028]  X is a bond or —O—;

[0029] A¹ is the same as A;

[0030] K is selected from the group consisting of N(R¹⁰)Y,

[0031]  and SO₂N(R⁸)(R¹⁰);

[0032]  D is a fused aryl or heteroaryl group;

[0033]  R¹¹ is selected from the group consising of alkoxy, aryloxy, andNHR¹²;

[0034]  R¹² is selected from the group consisting of H, alkyl, aryl, andheteroaryl, said alkyl, aryl or heteroaryl groups being optionallysubstituted with one or more J groups;

[0035]  Y is selected from the group consisting of SO₂R⁸, C(═O)NHR⁹,C(═S)NHR⁹, C(═NCN)R¹¹, C(═NC(═O)NHR¹⁰)R¹¹, and CO₂R⁸;

[0036]  R⁸ is selected from the group consisting of alkyl, alkoxy, aryl,and heterocyclyl, said alkyl, alkoxy, aryl, or heterocyclyl groups beingoptionally substituted with one or more J groups;

[0037]  R⁹ is selected from the group consisting of H, alkyl, aryl, andheteroaryl, said alkyl, aryl, or heteroaryl groups being optionallysubstituted with one or more J groups;

[0038]  or an R⁹ alkyl group may be combined with an A: alkylene groupto form a N-containing heterocyclic 5- or 6-membered ring;

[0039]  R¹⁰ is selected from the group consisting of H and lower alkyl;

[0040]  or in the moiety SO₂N(R⁸)R¹⁰, R⁸ and R¹⁰ may be combinedtogether with the N atom to which they are attached to form aN-containing heterocyclic 5- or 6-membered ring;

[0041]  or where A¹ is an alkylene group, and K is N(R¹⁰)Y wherein R¹⁰is alkyl, said R¹⁰ alkyl group may be combined with said A¹ alkylenegroup to form a N-containing heterocyclic 5- or 6-membered ring;

[0042]  or a pharmaceutically acceptable salt thereof.

[0043] In some preferred embodiments of the compounds of Formula I, nand v are each 0, q is 1, B is a bond, and G is H. In further preferredembodiments of the compounds of Formula I, R¹ is the sidechain of anaturally occurring amino acid. In still further preferred embodimentsof the compounds of Formula I, Ris —S(═C)R.

[0044] In some preferred embodiments of the compounds of Formula I, R²is benzyl or alkoxyalkyl. In more preferred embodiments, X is a bond,and Y is SO₂R⁸. Preferably, A¹ is —CH₂—CH₂—, —CH₂—CH(CH₃)—, or—(CH₃)CH—CH₂—.

[0045] In further preferred embodiments of the compounds of Formula I,R¹ is a serine sidechain, which is optionally capped with a benzylgroup. Preferably, the carbon to which the serine sidechain is attached,designated “M” in Formula I, is a carbon atom in the D configuration.

[0046] In preferred embodiments of the compounds of Formula I, R² isbenzyl, R⁷ is methyl, and R⁸ is substituted phenyl, unsubstitutedphenyl, substituted heteroaryl, or unsubstituted heteroaryl. Inparticularly preferred embodiments, R⁸ is aryl, aryl substituted withamino, aryl substituted with heterocyclomethyl, heteroaryl, alkylsubstituted with heteroaryl, or heteroaryl substituted with alkylthio,haloalkyl, alkyl, phenylsulfonyl, halogen, aminophenyl, amino, ordialkylaminoalkyl.

[0047] In further preferred embodiments of the compounds of Formula I,n, v and q are each 0, B is (C═O), and G is phenyl or lower alkyl, saidphenyl or lower alkyl groups being optionally substituted with one ormore J groups.

[0048] In more preferred embodiments of the invention, n and v are each0, q is 1, R¹ is the side chain of an amino acid in the D- orL-configuration, R³ is S(═O)₂R⁷, G is H, B is a bond, R² is benzyl oralkoxyalkyl, X is a bond, and Y is SO₂R.

[0049] In other preferred embodiments, A¹ is CH₂CH₂, CH₂CH(CH₃), or(CH₃)CHCH₂. In more preferred embodiments, R¹ is a serine side chain inthe D-configuration in which the hydroxyl group is capped with benzyl,R² is benzyl, R⁷ is methyl, and R⁸ is substituted or unsubstitutedphenyl or substituted or unsubstituted heteroaryl.

[0050] More preferred are the substituents shown for R₁-R₄, B, G, Aaa,X, A, Y, n, q and v shown for the compounds in Tables 2, 3, 4 and 5.Especially preferred are the substituents shown for compounds 9, 13, 17,22, and 29-151.

[0051] Some especially preferred embodiments of the compounds of FormulaI are shown in Tables 2, 3, 4 and 5, infra, with compounds 9, 13, 17,22, and 29-151 being particularly preferred.

[0052] Because the peptide-containing α-ketoamides of the inventioninhibit cysteine proteases and serine proteases, they can be used inboth research and therapeutic settings.

[0053] In a research environment, preferred compounds having definedattributes can be used to screen for natural and synthetic compoundswhich evidence similar characteristics in inhibiting protease activity.The compounds can also be used in the refinement of in vitro and in vivomodels for determining the effects of inhibition of particular proteaseson particular cell types or biological conditions.

[0054] In a therapeutic setting, given the connection between cysteineproteases and certain defined disorders, and serine proteases andcertain defined disorders, compounds of the invention can be utilized toalleviate, mediate, reduce and/or prevent disorders which are associatedwith abnormal and/or aberrant activity of cysteine proteases and/orserine proteases.

[0055] In preferred embodiments, compositions are provided forinhibiting a serine protease or a cysteine protease comprising acompound of the invention and a pharmaceutically acceptable carrier. Inother preferred embodiments, methods are provided for inhibiting serineproteases or cysteine proteases comprising contacting a proteaseselected from the group consisting of serine proteases and cysteineproteases with an inhibitory amount of a compound of the invention.

[0056] Methodologies for making the present peptide-containingα-ketoamide inhibitors are also disclosed. Other useful methodologieswill be apparent to those skilled in the art, once armed with thepresent disclosure. These and other features of the compounds of thesubject invention are set forth in more detail below.

DETAILED DESCRIPTION

[0057] Disclosed herein are the selected peptide-containing α-ketoamideswhich are represented by the following formula I:

[0058] wherein:

[0059] Q has the formula G-B—(CHR⁴)_(v) where R⁴ is independently H oralkyl having from 1 to 4 carbons;

[0060] v is 0, 1, or 2;

[0061] B is selected from the group consisting of C(═O), OC(═O),S(═O)_(m), CH₂, a bond, NR⁵C(═O), S(═O)_(m)-A-C(═O), and C(═O)-A-C(═O),where R⁵ is H or lower alkyl;

[0062] m is 0, 1, or 2;

[0063] A is lower alkylene or cycloalkylene, optionally substituted withone or more halogen atoms, aryl, or heteroaryl groups;

[0064] M is a carbon atom;

[0065] G is selected from the group consisting of H, a blocking group,lower alkyl, lower alkenyl, aryl having from about 6 to about 14carbons, heterocyclyl having from about 5 to about 14 ring atoms,heterocycloalkyl having from about 5 to about 14 ring atoms, arylalkylhaving from about 7 to about 15 carbons, heteroarylalkyl, andarylheteroalkyl wherein the aryl portion can be unfused or fused withthe heteroalkyl ring, said alkyl, aryl, heterocyclyl, heterocycloalkyl,arylalkyl, heteroarylalkyl, and arylheteroalkyl groups being optionallysubstituted with one or more J groups;

[0066] J is selected from the group consisting of halogen, CN, nitro,lower alkyl, cycloalkyl, heterocycloalkyl, heteroalkyl, halogenatedalkyl, aryloxyalkyl, alkylthio, alkylsulfonyl, aryl, heteroaryl,arylalkyl, arylalkyloxy, arylsulfonyl, heteroarylsulfonyl,alkoxycarbonyl, alkoxyalkyl, acyl, alkoxy, hydroxy, carboxy,hydroxyalkyl, amino, alkylamino, and aminoalkyl, said amino group orsaid amino group of said aminoalkyl or alkylamino group being optionallysubstituted with an acyl group, an alkoxy group, or with 1 to 3 aryl,lower alkyl, cycloalkyl, or alkoxyalkyl groups; and said aryl,heteroaryl, heterocycloalkyl, and heteroalkyl groups being furtheroptionally substituted by a J group;

[0067] each Aaa is independently an amino acid which optionally containsone or more blocking groups;

[0068] n is 0, 1, 2, or 3;

[0069] R¹ and R² are independently selected from the group consisting ofH, alkyl having from one to about 6 carbons, arylalkyl having from about7 to about 15 carbons, heteroalkyl in which the ring contains from about5 to about 14 ring atoms, heteroarylalkyl in which the heteroaryl ringcontains from about 5 to about 14 ring atoms, alkoxyalkyl, a side chainof a naturally occurring amino acid in the R or S configuration, and(CH₂)_(p)NH-L, said alkyl, arylalkyl, heteroalkyl, heteroarylalkyl, andalkoxyalkyl groups being optionally substituted with one or more Jgroups;

[0070] p is 0, 1, 2, or 3;

[0071] L is selected from the group consisting of alkoxycarbonyl havingfrom 2 to about 7 carbons, arylalkoxycarbonyl in which the arylalkoxygroup contains about 7 to about 15 carbons, and S(═O)R⁶;

[0072] R⁶ is selected from the group consisting of lower alkyl, and arylhaving from about 6 to about 14 carbons;

[0073] R³ is selected from the group consisting of H, alkyl having fromone to about 6 carbons, arylalkyl having from about 7 to about 15carbons, heteroalkyl in which the ring contains from about 5 to about 14ring atoms, heteroarylalkyl in which the heteroaryl ring contains fromabout 5 to about 14 ring atoms, alkoxyalkyl, a side chain of a naturallyoccurring amino acid in the R or S configuration, (CH₂)_(p)NH-L,C(═O)R⁷, S(═O)₂R⁷, a blocking group, and when combined with the carbonatom to which R¹ is attached an alkylene group having from 2 to 5carbons, said alkylene group being optionally substituted with agroup-selected from the group consisting of aryl, azide, CN, a protectedamino group, and OSO₂-aryl, said alkyl, arylalkyl, heteroalkyl,heteroarylalkyl, and alkoxyalkyl groups being optionally substitutedwith one or more J groups;

[0074] R⁷ is selected from the group consisting of aryl having fromabout 6 to about 14 carbons, heteroaryl having from about 5 to about 14ring atoms, arylalkyl having from about 7 to about 15 carbons, alkylhaving from 1 to about 10 carbons, said aryl, heteroaryl, arylalkyl andalkyl groups being optionally substituted with one or more J groups,heteroalkyl having from 2 to about 7 carbons, alkoxy having from about 1to about 10 carbons, and amino optionally substituted with 1 or morealkyl groups;

[0075] q is 0 or 1;

[0076] Z is selected from the group consisting of C(═O)C(═O)NH—X-A¹-Kand

[0077]  X is a bond or —O—;

[0078]  A¹ is the same as A;

[0079]  K is selected from the group consisting of N(R¹⁰)Y,

[0080]  and SO₂N(R⁸)(R¹⁰);

[0081]  D is a fused aryl or heteroaryl group;

[0082]  R¹¹ is selected from the group consising of alkoxy, aryloxy, andNHR¹²;

[0083]  R¹² is selected from the group consisting of H, alkyl, aryl, andheteroaryl, said alkyl, aryl or heteroaryl groups being optionallysubstituted with one or more J groups;

[0084]  Y is selected from the group consisting of SOR⁸, C(═O)NHR⁹,C(═S)NHR⁹, C(═NCN)R¹¹, C(═NC(═O)NHR¹⁰)R¹¹, and COR;

[0085]  R⁸ is selected from the group consisting of alkyl, alkoxy, aryl,and heterocyclyl, said alkyl, alkoxy, aryl, or heterocyclyl groups beingoptionally substituted with one or more J groups;

[0086]  R⁹ is selected from the group consisting of H, alkyl, aryl, andheteroaryl, said alkyl, aryl, or heteroaryl groups being optionallysubstituted with one or more J groups;

[0087]  or an R⁹ alkyl group may be combined with an A¹ alkylene groupto form a N-containing heterocyclic 5- or 6-membered ring;

[0088]  R¹⁰ is selected from the group consisting of H and lower alkyl;

[0089]  or in the moiety SO₂N(R⁸)R¹⁰, R⁸ and R¹⁰ may be combinedtogether with the N atom to which they are attached to form aN-containing heterocyclic 5- or 6-membered ring;

[0090]  or where A¹ is an alkylene group, and K is N(R¹⁰)Y wherein R¹⁰is alkyl, said R¹⁰ alkyl group may be combined with said A¹ alkylenegroup to form a N-containing heterocyclic 5- or 6-membered ring;

[0091]  or a pharmaceutically acceptable salt thereof.

[0092] It is recognized that various stereoisomeric forms of thecompounds of Formula I may exist. Preferred compounds of the inventionhave any Aaa groups being α-amino acids in the L-configuration. However,racemates and individual enantiomers and mixtures thereof form part ofthe present invention.

[0093] The carbon atom designated as “M” in the compounds of Formula Ican exist in either the D or the L configuration. In some preferredembodiments, M is a carbon atom having the “D” configuration.

[0094] As used herein, the term “alkyl” includes straight-chain, andbranched hydrocarbon groups such as, for example, methyl, ethyl, propyl,isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl,1-ethylpentyl, hexyl, and octyl groups. “Cycloalkyl” groups are cyclicalkyl groups, such as, for example, cyclopropyl, methylcyclopentyl, andcyclohexyl groups. Preferred alkyl groups have 1 to about 10 carbonatoms, most preferably “lower alkyl” of 1 to about 6 carbon atoms.“Alkylene” groups are alkyl groups having two points of attachment;i.e., non-terminal alkyl groups. “Lower alkylene” groups are branched orunbranched alkylene groups of 1 to about 6 carbon atoms such as, forexample, ethylene(—CH₂CH₂—), propylene, butylene, hexylene,1-methylethylene, 2-methylethylene, and 2-methylpropylene.“Cycloalkylene” groups are cyclic alkylene groups. “Acyl” groups arealkylcarbonyl groups. “Aryl” groups are aromatic cyclic compoundspreferably including but not limited to phenyl, tolyl, naphthyl,anthracyl, phenanthryl, and pyrenyl. Also included within the definitionof “aryl” are ring systems having two aromatic rings connected by abond, such as biphenyl. Preferred aryl groups include phenyl andnaphthyl.

[0095] The term “carbocyclic”, as used herein, refers to cyclic groupsin which the ring portion is composed solely of carbon atoms. The term“halogen” refers to F, Cl, Br, and I atoms. The term “arylalkyl” denotesalkyl groups which bear aryl groups, for example, benzyl groups. As usedherein, “alkoxy” groups are alkyl groups linked through an oxygen atom.Examples of alkoxy groups include methoxy (—OCH₃) and ethoxy (—OCH₂CH₃)groups. In general, the term “oxy” when used as a suffix denotesattachment through an oxygen atom. Thus, alkoxycarbonyl groups arecarbonyl groups which contain an alkoxy substituent, i.e., groups ofgeneral formula —C(═O)—O—R, where R is alkyl. The term “alkoxyalkyl”denotes an alkoxy group attached to an alkyl group. The term “aryloxy”denotes an aryl group linked through an oxygen atom, and the term“arylalkyloxy” denotes an arylalkyl group linked through an oxygen atom.

[0096] The terms “heterocycle”, “heterocyclyl”, and “heterocyclic” referto cyclic groups in which a ring portion includes at least oneheteroatom such as O, N or S. Heterocyclic groups include “heteroaryl”as well as “heteroalkyl” groups. The term “heteroaryl” denotes arylgroups having one or more hetero atoms (e.g., O, N, or S) containedwithin an aromatic ring. Also included within the definition of“heteroaryl” are ring systems having two aromatic rings connected by abond, where at least one of the rings contains a hetero atom. Preferred“heteroaryl” groups include pyridyl, pyrimidyl, pyrrolyl, furyl,thienyl, imidazolyl, triazolyl, tetrazolyl, quinolyl, isoquinolyl,benzoimidazolyl, thiazolyl, bipyridyl, pyridylthiophenyl,pyrimidylthiophenyl, benzimidazolyl, isoxazolylthiophenyl,pyrazolylthiophenyl, phthalimido, and benzothiazolyl. The term“heterocycloalkyl” denotes a heterocycle attached through a lower alkylgroup. The term “heteroarylalkyl” denotes a heteroaryl group attachedthrough an alkyl group. As used herein, the term “heteroalkyl” denotes aheterocyclic group which contains at least one saturated carbon atom ina heterocyclic ring. Examples of heteroalkyl groups include piperidine,dihydropyridine, and tetrahydroisoquinyl groups. The term“arylheteroalkyl” as used herein denotes a“heteroalkyl” group connectedthrough an aryl group. One preferred example of an arylheteroalkyl groupis dibenzo-γ-pyranyl.

[0097] As used herein, the term “amino acid” denotes a moleculecontaining both an amino group and a carboxyl group. As used herein theterm “L-amino acid” denotes an α-amino acid having the L-configurationaround the α-carbon, that is, a carboxylic acid of general formulaCH(COOH)(NH₂)— (side chain), having the L-configuration. The term“D-amino acid” similarly denotes a carboxylic acid of general formulaCH(COOH)(NH₂)— (side chain), having the D-configuration around theα-carbon. Side chains of L-amino acids include naturally occurring andnon-naturally occurring moieties. Nonnaturally occurring (i.e.,unnatural), amino acid side chains are moieties that are used in placeof naturally occurring amino acid sidechains in, for example, amino acidanalogs. See, for example, Lehninger, Biochemistry, Second Edition,Worth Publishers, Inc, 1975, pages 73-75. One representative amino acidside chain is the lysyl side chain, —(CH₂)₄—NH₂. Other representativeα-amino acid side chains are shown below in Table 1. TABLE 1 CH₃—HS—CH₂— HO—CH₂— HO₂C—CH(NH₂)—CH₂—S—S—CH₂— C₆H₅—CH₂— CH₃—CH₂—HO—C₆H₄—CH₂— CH₃—S—CH₂—CH₂—

CH₃—CH₂—S—CH₂—CH₂—HO—CH₂—CH₂—CH₃—CH(OH)—HO₂C—CH₂—NHC(═O)—CH₂—

HO₂C—CH₂—CH₂—NH₂C(═O)—CH₂—CH₂—(CH₃)₂—CH—(CH₃)₂—CH—CH₂—CH₃—CH₂—CH₂—H₂N—CH₂—CH₂—CH₂—

H₂N—C(═NH)—NH—CH₂—CH₂—CH₂—H₂N—C(═O)—CH₂—CH₂—CH₂—CH₃—CH₂—CH(CH₃)—CH₃—CH₂—CH₂—CH₂—H₂N—CH₂—CH₂—CH₂—CH₂—

[0098] Functional groups present in the compounds of Formula I maycontain blocking groups. Blocking groups are known per se as chemicalfunctional groups that can be selectively appended to functionalities,such as hydroxyl groups, amino groups, thio groups, and carboxyl groups.Protecting groups are blocking groups which can be readily removed fromfunctionalities. These groups are present in a chemical compound torender such functionality inert to chemical reaction conditions to whichthe compound is exposed. Any of a variety of protecting groups may beemployed with the present invention. Examples of such protecting groupsare the benzyloxycarbonyl (Cbz; Z), toluenesulfonyl, t-butoxycarbonyl,methyl ester, and benzyl ether groups. Other preferred protecting groupsaccording to the invention may be found in Greene, T. W. and Wuts, P. G.M., “Protective Groups in Organic Synthesis” 2d. Ed., Wiley & Sons,1991, which is hereby incorporated by reference in its entirety.

[0099] Further blocking groups useful in the compounds of the presentinvention include those that bear acyl, aroyl, alkyl, alkanesulfonyl,arylalkanesulfonyl, or arylsulfonyl substituents on their amino groups.Other useful blocking groups include alkyl ethers, e.g., the methylether of serine.

[0100] The disclosed compounds of the invention are useful for theinhibition of cysteine proteases and serine proteases. As used herein,the terms “inhibit” and “inhibition” mean having an adverse effect onenzymatic activity. An inhibitory amount is an amount of a compound ofthe invention effective to inhibit a cysteine and/or serine protease.

[0101] Pharmaceutically acceptable salts of the cysteine and serineprotease inhibitors also fall within the scope of the compounds asdisclosed herein. The term “pharmaceutically acceptable salts” as usedherein means an inorganic acid addition salt such as hydrochloride,sulfate, and phosphate, or an organic acid addition salt such asacetate, maleate, fumarate, tartrate, and citrate. Examples ofpharmaceutically acceptable metal salts are alkali metal salts such assodium salt and potassium salt, alkaline earth metal salts such asmagnesium salt and calcium salt, aluminum salt, and zinc salt. Examplesof pharmaceutically acceptable organic amine addition salts are saltswith morpholine and piperidine. Examples of pharmaceutically acceptableamino acid addition salts are salts with lysine, glycine, andphenylalanine.

[0102] Compounds provided herein can be formulated into pharmaceuticalcompositions by admixture with pharmaceutically acceptable nontoxicexcipients and carriers. As noted above, such compositions may beprepared for use in parenteral administration, particularly in the formof liquid solutions or suspensions; or oral administration, particularlyin the form of tablets or capsules; or intranasally, particularly in theform of powders, nasal drops, or aerosols; or dermally, via, forexample, transdermal patches; or prepared in other suitable fashions forthese and other forms of administration as will be apparent to thoseskilled in the art.

[0103] The composition may conveniently be administered in unit dosageform and may be prepared by any of the methods well known in thepharmaceutical art, for example, as described in Remington'sPharmaceutical Sciences (Mack Pub. Co., Easton, Pa., 1980). Formulationsfor parenteral administration may contain as common excipients sterilewater or saline, polyalkylene glycols such as polyethylene glycol, oilsand vegetable origin, hydrogenated naphthalenes and the like. Inparticular, biocompatible, biodegradable lactide polymer,lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylenecopolymers may be useful excipients to control the release of the activecompounds. Other potentially useful parenteral delivery systems forthese active compounds include ethylene-vinyl acetate copolymerparticles, osmotic pumps, implantable infusion systems, cyclodextrinsand liposomes. Formulations for inhalation administration contain asexcipients, for example, lactose, or may be aqueous solutionscontaining, for example, polyoxyethylene-9-lauryl ether, glycocholateand deoxycholate, or oily solutions for administration in the form ofnasal drops, or as a gel to be applied intranasally. Formulations forparenteral administration may also include glycocholate for buccaladministration, a salicylate for rectal administration, or citric acidfor vaginal administration. Formulations for transdermal patches arepreferably lipophilic emulsions.

[0104] The materials for this invention can be employed as the soleactive agent in a pharmaceutical or can be used in combination withother active ingredients which could facilitate inhibition of cysteineand serine proteases in diseases or disorders.

[0105] The concentrations of the compounds described herein in atherapeutic composition will vary depending upon a number of factors,including the dosage of the drug to be administered, the chemicalcharacteristics (e.g., hydrophobicity) of the compounds employed, andthe route of administration. In general terms, the compounds of thisinvention may be provided in effective inhibitory amounts in an aqueousphysiological buffer solution containing about 0.1 to 10% w/v compoundfor parenteral administration. Typical dose ranges are from about 1μg/kg to about 1 g/kg of body weight per day; a preferred dose range isfrom about 0.01 mg/kg to 100 mg/kg of body weight per day. Suchformulations typically provide inhibitory amounts of the compound of theinvention. The preferred dosage of drug to be administered is likely,however, to depend on such variables as the type or extent ofprogression of the disease or disorder, the overall health status of theparticular patient, the relative biological efficacy of the compoundselected, and formulation of the compound excipient, and its route ofadministration.

[0106] As used herein, the term “contacting” means directly orindirectly causing at least two moieties to come into physicalassociation with each other. Contacting thus includes physical acts suchas placing the moieties together in a container, or administeringmoieties to a patient. Thus, for example administering a compound of theinvention to a human patient evidencing a disease or disorder associatedwith abnormal and/or aberrant activity of such proteases falls withinthe scope of the definition of the term “contacting”.

[0107] The invention is further illustrated by way of the followingexamples which are intended to elucidate the invention. These examplesare not intended, nor are they to be construed, as limiting the scope ofthe disclosure.

EXAMPLES

[0108] General Methods. Thin layer chromatography was performed onsilica gel plates (MK6F 60A, size 1×3 in, layer thickness 250 μm,Whatman Inc.). Preparative thin layer chromatography was performed onsilica gel plates (size 20×20 in, layer thickness 1000 micron,Analtech). Preparative column chromatography was carried out using Mercksilica gel, 40-63 μm, 230-400 mesh. ¹H NMR spectra were recorded on a GEQE Plus instrument (300 MHZ) using tetramethylsilane as internalstandard. Electrospray mass spectra were recorded on a VG platform IIinstrument (Fisons Instruments).

[0109] Compounds of the invention were prepared following one of theGeneral Methods A, B, C or D.

Example 1 Preparation of Compound 9 by General Method A

[0110]

Preparation of Compound 1

[0111] This compound and related hydroxy acids used in this study weresynthesized following a general procedure of Harbeson et al, J. Med.Chem. 1994, 37, 2918-2929, which is incorporated herein by reference inits entirety.

Preparation of Compound 2

[0112] To a cooled (−10 ° C.) solution of compound 1 (4.30 g, 0.015 mol)in anhydrous methanol (50 mL) was added slowly thionyl chloride (3.20mL). After 0.5 hour, the cooling bath was removed, the mixture wasstirred for an additional 16 hours and concentrated to give a residuewhich on trituration with ethyl acetate (30 mL) gave a white solid. Thesolid was separated by filtration and dried to give 3.50 g of compound 2which was used directly in the next step; MS m/e 210(M+H).

Preparation of Compound 3

[0113] The preparation of this compound is shown in General Method E.

Preparation of Compound 4

[0114] To a cooled (0° C.) solution of compound 1 (1.00 g, 0.0034 mol)in anhydrous DMF (20 mL) was added NMM (1.40 g, 0.014 mmol) followed by1-HOBt (0.54 g, 0.0040 mmol) and BOP (1.80 g, 0.0040 mmol). The mixturewas stirred for 15 minutes and to it was added compound 3 (0.75 g,0.0032 mmol). The cooling bath was removed and the mixture was stirredfor 4 hours, poured into ice-water (200 mL), and extracted into ethylacetate (3×100 mL). The organic layer was washed with 2% citric acidsolution (2×50 mL), 2% sodium bicarbonate solution (2×50 mL), brine(1×50 mL), and it was dried over anhydrous sodium sulfate. Solventevaporation under reduced pressure gave a crude solid which was washedseveral times with n-pentane to produce 1.30 g of compound 4.

[0115] Compound 4: white solid (diastereomeric mixture); ¹H-NMR(DMSO-d₆) δ 7.90 and 7.65 (2 sets of t, 1H), 7.75 (d, 2H), 7.55 (q, 2H),7.15 (m, 6H), 6.55 and 5.80 (2 sets of d, 1H), 3.90 (m, 2H), 3.30 (d,1H), 3.10 (m, 2H), 2.75 (m, 2H), 2.50 (m, 3H), 1.20 (s, 9H). MS m/e478(M+H), 500(M+Na).

Preparation of Compound 5

[0116] To a solution of compound 4 (0.40 g, 0.84 mmol) in 1,4-dioxane(15 mL) was added 4 N HCl in dioxane (15 mL). The reaction mixture wasstirred at room temperature for 2 hours, then concentrated at reducedpressure to give a residue which was washed several times with ethylacetate and dried under vacuum to give 0.30 g of compound 5; ¹H-NMR(DMSO-d₆) showed complete absence of tBoc peak at δ 1.20 ppm; MS m/e 378(M+H). This material was used directly in the next step.

Preparation of Compound 7

[0117] To a mixture of D-Ser(Bn) (compound 6, 1.00g, 5 mmol) and 1 NNaOH (10 mL, 10 mmol) at 0° C. was slowly added methanesulfonyl chloride(0.80 g, 7.69 mmol). After 0.5 hour, the cooling bath was removed, themixture was stirred overnight and acidified (pH˜2-3) with 2 N HCl. Theaqueous layer was extracted into ethyl acetate (3×50 ML). The combinedorganic layer was washed with water (1×20 mL) and brine (1×20 mL), anddried over MgSO₄. Solvent evaporation gave a residue which wasredissolved in methylene chloride (10 mL); addition of hexanes produceda white solid which was filtered and dried to give 1.02 g of compound 7.

Preparation of Compound 8

[0118] This compound was prepared by coupling compound 7 and compound 5,using NMM/HOBt/BOP as coupling agents, following the procedure describedabove for the preparation of compound 4. In some of the relatedexamples, EDCI/HOBt were used as coupling agents.

Preparation of Compound 9

[0119] To a cooled (0° C.) solution of compound 8 (0.31 g, 0.49 mmol) inanhydrous methylene chloride (10 mL) was added Dess-Martin periodinanereagent (0.425 g, 1.00 mmol). The cooling bath was removed and themixture was stirred for an additional 1 hour. The solution was thendiluted with methylene chloride (10 mL), and washed with 10% sodiumthiosulfate solution (5×5 mL), saturated sodium bicarbonate solution(2×5 mL), and brine (1×5 mL), and dried over anhydrous sodium sulfate.Solvent removal under reduced pressure gave a residue which was washedwith n-pentane (10 mL) and dried under vacuum to produce 0.178 g ofcompound 9; ¹H-NMR spectrum revealed a minor amount of epimerization hadtaken place.

[0120] Compound 9: white solid; ¹H-NMR (DMSO-d₆) δ 8.75 (t, 1H), 8.60and 8.50 (2 doublets, 1H), 7.75 (d, 2H), 7.65-7.00 (a series of m, 15H),5.25 (broad m, 1H), 4.45 and 4.235 (2 singlets, 2H), 4.15 (m, 1H),3.35-2.60 (a series of m, 8H), 3.35 and 3.25 (2 singlets, 3H) MS m/e631(M+H), 653(M+Na).

Example 2 Preparation of Compound 13 by General Method B

[0121]

[0122] In General Method B, compound 4, prepared as described above, wasoxidized by Dess-Martin periodinane reagent to generate compound 10which on tBoc-deprotection (2 N HCl in dioxane) produced the amine-salt,compound 11. Coupling (NMM/HOBt/BOP) of compound 11 withN-phenylsulfonyl-(L)-Pro (compound 12) yielded compound 13. Purificationwas achieved by passing a solution of the crude material in methylenechloride through Sep-Pak® Vac 6 cc (1 g) silica cartidge (WatersCorporation, Milford, Mass.), eluting with methylene chloride, followedby various combinations of methylene chloride and ethyl acetate.Harbeson et al. (J. Med. Chem. 1994, 37, 2918-2929) reported that silicagel chromatography of a ketoamide epimerizes the chiral center at P₁.

[0123] Compound 13: white solid; ¹H-NMR (CDCl₃) δ 7.90-7.00 (a series ofm, 18H), 5.40 and 5.30 (2 multiplets, 1H), 4.10 (m, 1H), 3.50-3.00 (m,8H), 1.90-1.40 (m, 4H). MS m/e 613(M+H), 635(M+Na).

Example 3 Preparation of Compound 17 by General Method C

[0124] In General Method C, compound 2 was coupled (NMM/HOBt/BOP) withL-Cbz-Leu to give compound 14 which was hydrolyzed (aq. NaOH) tocompound 15. Coupling (NMH/HOBt/BOP) of compound 15 with compound 3 gavecompound 16 which underwent Dess-Martin oxidation to generate compound17.

[0125] Compound 17: white solid; ¹H-NMR (CDCl₃) δ 7.85 (d, 2H),7.60-7.00 (a series of m, 15H), 6.60 (d, 1H), 5.40 (m, 1H), 5.20 (q,1H), 5.10 (s, 2H), 4.10 (broad, 1H), 3.50-3.00 (a series of m, 6H),1.65-1.30 (m, 3H), 0.90 (d, 6H). MS m/e 623(M+H), 645(M+Na).

Example 4 Preparation of Compound 22 by General Method D

[0126] In General Method D, compound 7 was coupled (NMM/HOBt/BOP) withcompound 2 to generate compound 18 which underwent Dess-Martin oxidationto generate compound 19. Hydrolysis (LiOH, MeOH-H₂O) of compound 19 gavecompound 20 which was coupled (NMM/HOBt/BOP) with compound 21 to givecompound 22. Compound 22 was purified by silica gel chromatography.

[0127] Compound 22: white solid; MS m/e 646(M+H), 668(M+Na).

Preparation of Intermediates Example 5

[0128] The preparation of a representative example of an amine (compound3), containing a terminal sulfonamide moiety, is shown in General MethodE.

Preparation of Compound 24

[0129] To a solution of 1,2-ethylenediamine (compound 23, 10.80 g, 12.00mL, 0.18 mol) in THF (30 mL) was added slowly BOC—ON (22.10 g, 0.09 mol)in THF (70 mL) over a period of 4 hours. The reaction mixture wasstirred overnight, concentrated on a rotavapor, and taken up into water(150 mL). The aqueous layer was acidified (pH˜5-6) with solid citricacid monohydrate, washed with ether (3×50 mL) and then treated (at 0°C.) with 6 N NaOH solution to make it basic (pH˜12-13). The basicsolution was extracted into ethyl acetate (3×100 mL), and the combinedethyl acetate layer was dried (MgSO₄) and concentrated to generate 7.23g of monoprotected diamine, compound 24.

[0130] Compound 24: viscous liquid; ¹H-NMR (CDCl₃) δ 5.00 (broad, 1H),3.20 (broad q, 2H), 2.80 (t, 2H), 1.45 (s, 9H), 1.25 (broad, 2H).

Preparation of Compound 25

[0131] A cooled (0-5° C.) solution of compound 24 (0.321 g, 0.002 mol)in methylene chloride (5 mL) was treated sequentially with triethylamine(0.243 g, 0.33 mL, 0.0024 mol) and benzenesulfonyl chloride (0.423 g,0.30 mL, 0.0024 mol). The ice-bath was removed and the mixture wasstirred for an additional 0.5 hour, washed successively with water (2×5mL), cold (0-5° C.) 0.5 N HCl (1×5 mL, 2% NaHCO₃ solution (1×5 mL), andbrine (1×5 mL). The solution was dried (MgSO₄) and the solvent wasevaporated to give a residue which was washed several times withn-pentane. A total of 0.60 g of the sulfonamide derivative, compound 25,was obtained.

[0132] Compound 25: white solid, mp 92-95° C.; R_(f) (TLC, 5% methanolin methylene chloride) 0.55; ¹H-NMR (CDCl₃) δ 7.85 (d, 2H), 7.55 (m,3H), 5.30 (broad d, 1H), 4.85 (broad, 1H), 3.25 (broad q, 2H), 3.10(broad q, 2H), 1.40 (s, 9H).

Preparation of Compound 3

[0133] A solution of compound 25 (0.560 g, 0.0019 mol) in 1,4-dioxane (4mL) was treated with 4 N HCl in dioxane (4 mL). The mixture was stirredat room temperature for 1 hour and concentrated at the rotavapor. Theresidue was washed several times with ethyl acetate and dried undervacuum to give 0.40 g of compound 3.

[0134] Compound 3: white solid, mp 178-180° C.; ¹H-NMR (DMSO-d₆) δ8.20-8.00 (broad t, 4H), 7.80 (d, 2H), 7.60 (m, 3H), 2.95 (broad q, 2H),2.80 (broad, 2H).

Example 6 Preparation of Compound 28

[0135] The preparation of a representative example of an intermediateamine (compound 28) containing a terminal biaryl sulfonamide moiety isshown in General Method F.

[0136] A mixture of compound 26 (prepared from compound 24 and5-bromothiophene-2-sulfonyl chloride, following the same generalprocedure as described above for the preparation of compound 25, 0.50 g,1 eqv), dimethoxyethane (10 mL), 2 M Na₂CO₃ (5 eqv), phenylboronic acid(1.40 eqv) and Pd(PPh₃)₄ (0.04 eqv) was heated at 135° C. for 2.5 hours.The reaction mixture was concentrated at the rotavapor, and the residuewas taken up into water (20 mL). The aqueous layer was acidified withcitric acid and extracted into methylene chloride (3×20 mL). Thecombined organic layer was washed with water (1×10 mL) and brine (1×10mL). It was dried (MgSO₄) and concentrated to a small volume.Trituration of the residue with hexanes gave a solid which was separatedby filtration and dried under vacuum to produce 0.37 g of compound 27;¹H-NMR (CDCl₃) δ 7.60-7.20 (a series of m, 7H), 5.35 (broad, 1H), 4.85(broad, 1H), 3.30 (m, 2H), 3.20 (m, 2H), 1.40 (s, 9H). For a generaldescription of this reaction procedure, see Miyaura et al., Chem. Rev.1995, 95, 2457-2483.

[0137] Compound 27 was converted to compound 28 following the proceduredescribed for the preparation of compound 3.

Example 7 Preparation of Taurine Sulfonamide Intermediate

[0138] The preparation of a representative taurine sulfonamideintermediate is shown in General Method G.

[0139] The phthalimide of taurine, prepared by a known procedure (R.Winterbottom et al., J. Amer. Chem. Soc., 1947, 69, 1393-1401) wasconverted to its sulfonyl chloride with phosphorous pentachloride inrefluxing benzene. This was allowed to react with aniline in thepresence of pyridine to form the corresponding sulfonamide. Thephthalimide protecting group was then removed by refluxing withhydrazine and the resulting taurine sulfonamide was isolated as itshydrochloride.

Example 8

[0140] Syntheses of compounds 29 through 50 in Tables 2 and 3 werecarried out using the designated general methods, as described, and theappropriate starting materials.

Example 9 Inhibition of Cysteine Protease Activity

[0141] To evaluate inhibitory activity, stock solutions (40 timesconcentrated) of each compound to be tested were prepared in 100%anhydrous DMSO and 5 μl of each inhibitor preparation were aliquotedinto each of three wells of a 96-well plate. Recombinant human calpainI, prepared by the method of Meyer et al. (Biochem. J. 1996, 314:511-519; incorporated herein by reference in its entirety), was dilutedinto assay buffer (i.e., 50 mM Tris, 50 mM NaCl, 1 mM EDTA, 1 mM EGTA,and 5 mM β-mercaptoethanol, pH 7.5, including 0.2 mM Succ-Leu-Tyr-MNA),and 175 μl was aliquoted into the same wells containing the independentinhibitor stocks as well as to positive control wells containing 5 μlDMSO, but no compound. To start the reaction, 20 μl of 50 mM CaCl₂ inassay buffer was added to all wells of the plate, excepting three, whichwere used as background signal baseline controls. Substrate hydrolysiswas monitored every 5 minutes for a total of 30 minutes. Substratehydrolysis in the absence of inhibitor was linear for up to 15 minutes.

[0142] Inhibition of calpain I activity was calculated as the percentdecrease in the rate of substrate hydrolysis in the presence ofinhibitor relative to the rate in its absence. Comparison between theinhibited and control rates was made within the linear range forsubstrate hydrolysis. The IC₅₀s of inhibitors (concentration yielding50% inhibition) were determined from the percent decrease in rates ofsubstrate hydrolysis in the presence of five to seven differentconcentrations of the test compound. The results were plotted as percentinhibition versus log inhibitor concentration, and the IC₅0 wascalculated by fitting the data to the four-parameter logistic equationshown below using the program GraphPad Prism (GraphPad Software, Inc.,San Diego, Calif.).

y=d+[(a−d)/(1+(x/c)^(b))]

[0143] The parameters a, b, c, and d are defined as follows: a is %inhibition in the absence of inhibitor, b is the slope, c is the IC₅₀,and d is the % inhibition at an infinite concentration of inhibitor.

[0144] Results are presented in Tables 2, 3, 4 and 5 below. TABLE 2Inhibitory Activity of Linear α-Ketoamides

Cmp. Calpain Prep. MS No. W R₂ R IC₅₀ nM Method (M + 1)*  9 Ms-D- Bn Ph16 A 631 Ser(Bn) 13 PhSO₂-L- Bn Ph (78%) B 613 Pro ** 17 Cbz-L-Leu Bn Ph11 C 623 22 Ms-D- Ser(Bn) Bn

42 D 646 29 Ms-D- Ser(Bn) Bn

14 A 714 30 PhSO₂-L- Bn Ph (97%) B 663 Phe ** 31 Ms-D- Ser(Bn) Bn

29 A 632 32 Ms-D- Ser(Bn) Bn

10 A 704 33 Ms-D- Ser(Bn) Bn

17 A 761 34 Ms-D- Ser(Bn) Bn

25 A 786 (M + 2)* 35 Ms-D- Bn CH₃ 91 A 569 Ser(Bn) 36* Ms-D- Ser(Bn)CH₂OMe

14 A 668 37 Ms-D- Ser(Bn) Bn

18 C 777 38* Ms-D- CH₂OMe Ph  (100%) A 585 Ser(Bn) * 39 Ms-D- Ser(Bn) Bn

22 C 715, 777 ⁷⁹Br, ⁸¹Br 40 Ms-D- Ser(Bn) Bn

63 D 722 41 Ms-D- Ser(Bn) Bn

(88%) ** D 699 42 Ms-D- Ser(Bn) Bn

14 C 712(M)* 43 Ms-D- Ser(Bn) Bn

11 C 718(M)* 44 Ms-D- Ser(Bn) Bn

31 D 729 (M + 2)* 45 Ms-D- Ser(Bn) Bn

64 D 688 46 Ms-D- Ser(Bn) Bn

 144 D 660

[0145] TABLE 3 Inhibitory Activity of Branched-Chain α-Ketoamides

Ex. Calpain IC₅₀ Prep. No. D nM Method MS (M + 1) 47

26 A 645 48

43 A 645 49

32 A 646 (M + 2 50 CH₂CH₂N(CH₃)SO₂Ph (100%) C 645 **

[0146] Compounds listed in Table 4 were prepared by the general methodsA-G described above. TABLE 4 Inhibitory Activity of α-Ketoamides

Ex. Calpain MS No. W R IC₅₀ nM (M + 1) 51 Ms-D-SerCH₂CH₂NHSO₂-(3-(2-NH₂-thiazol-4-yl)Ph) 29 729 (Bn) 52 Ms-D-SerCH₂CH₂NHSO₂-(5-(3-formylphenyl)thiophene- 5 741 (Bn) 2-yl) 53 Ms-D-SerCH₂CH₂NHC(═N—CN)OPh 15 635 (Bn) 54 Ms-D-Ser CH₂CH₂NHSO₂(5-(3- 12 770(Bn) (Me₂NCH₂)phenyl)thiophene-2-yl) 55 Ms-D-Ser 3-Boc-NH-cyclohexane 42645 (Bn) 56 Ms-D-Ser CH₂CH₂NHSO₁(5-(3- 18 812 (Bn)(morpholinoCH₂)phenyl)thiophene-2-yl) 57 Ms-D-SerCH₂CH₂NHSO₂(4-(MorpholinoCH₂)Ph) 18 730 (Bn) 58 Ms-D-SerCH₂CH₂NHSO₂(5-(3-(N-Me-piperazinyl- 21 825 (Bn)CH₂)phenyl)thiophene-2-yl) 59 Ms-D-Ser CH₂CH₂NHSO₂(5-(3- 35 765 (Bn)(HOCH₂)phenyl)phenyl)thiophene-2-yl) (2 diast) (M + Na) 60 Ms-D-SerCH₂CH₂SO₂NHPh 47 631 (Bn) 61 Ms-D-Ser CH₂CH₂SO₂NH(4-CF₃Ph) 32 699 (Bn)62 Ms-D-Ser (CH₂)₃SO₂NHPh 18 645 (Bn) 63 Ms-D-Ser (CH₂)₃SO₂NH(4-CF₃Ph)23 713 (Bn) 64 Ms-D-Ser 6-ketopiperidin-3-yl (33)* 545 (Bn) 65 Ms-D-SerCH₂CH₂N(Me)SO₂-(5-(3- 19 755 (Bn) formylphenyl)thiophene-2-yl) 66Ms-D-Thr CH₂CH₂NHSO₂(5-pyrid-2-ylthiophene-2-yl) 12 728 (Bn) 67 Ms-D-Ser-N(Me)SO₂-(5-isoxazol-3-yl-thiophene-2-yl) 21 718 (Bn) 68 Ms-(D,L)-CH₂CH₂NHSO₂(5-pyrid-2-ylthiophene-2-yl) 21 670 Phenylgly 69 Ms-(D,L)-CH₂CH₂NHSO₂Ph 80 587 Phenylgly 70 Ms-D-Thr CH₂CH₂NHSO₂(5-(3- 23 826 (Bn)(morpholinoCH₂)phenyl)thiophene-2-yl) (Mixture of diasteromers) 71Ms-D-Phe CH₂CH₂NHSO₂(5-pyrid-2-ylthiophene-2-yl) 18 684 72 Ms-D-SerCH₂CH₂NHSO₂(5-(3-Fluorophenyl)thiophene-2- 18 731 (Bn) yl) (Mixture ofdiastereomers) 73 Ms-D-Ser (CH₂)₃NHOCH₃ 87 597 (Bn) (M − 1) 74 Ms-D-SerCH₂CH₂NHSO₂-(5-(3-Nitrophenyl)thiophene-2- 15 758 (Bn) yl) (Mixture ofdiastereomers) 75 Ms-D-Ser CH₂CH₂NHSO₂(5-(3-Methylphenyl)thiophene- 36727 (Bn) 2-yl) (Mixture of diastereomers) 76 Ms-D-SerCH₂CH₂NHSO₂(5-(3-(AcNH)phenyl)thiophene- 11 792 (Bn) 2-yl) (M + Na)(Mixture of diastereomers 77 Ms-D-Ser CH₂CH₂NHSO₂(5-(3- 10 777 (Bn)(CH₃CO)phenyl)thiophene-2-yl) (M + Na) (Mixture of diastereomers) 78Ms-D-Ser 1-(4- 48 770 (Bn) (Morpholinomethyl)benzenesulfonyl)piperidin-4-yl) (Mixture of diastereomers) 79 Ms-D-Ser CH₂CH₂NHSO₂((4- 11 847 (Bn)(CH₃COPh)piperazin-1-yl)CH₂Ph) (Mixture of diastereomers) 80 Ms-D-Ser(CH₂)₃SO₂NH-morpholin-4-yl 169 652 (Bn) (M − 1) 81 Ms-D-Ser(CH₂)₃SO₂-morpholin-4-y1 124 639 (Bn) 82 Ms-D-Ser CH₂CH₂NHSO₂-(5-(4- 13765 (Bn) Methoxyphenyl)thiophene-2-yl) (M + Na) (Mixture ofdiastereomers) 83 Ms-D-Ser CH₂CH₂CH₂-Saccharin 48 657 (Bn) 84 Ms-D-SerCH₂CH₂NHSO₂((4-(PhCH₂)piperazin-1- 23 819 (Bn) yl)CH₂Ph) 85 Ms-D-SerCH₂CH₂NHSO₂((4-(CH₃CO)piperazin-l- 14 771 (Bn) yl)CH₂Ph) 86 Ms-D-SerCH₂CH₂NHSO₂-(5-Me₂N-naphth-1-yl) 49 724 (Bn) 87 Ms-D-SerCH₂CH₂NHSO₂-benzothiophene-2-yl 23 687 (Bn) 88 Cbz-Leu-CH₂CH₂NHSO₂(5-pyrid-2-ylthiophene- 33 819 Leu 2-yl) 89 Ms-D-SerCH₂CH₂NHSO₂((4-Pyrid-2-yl)piperazin- 21 806 (Bn) 1-yl)CH₂Ph) (Mixture ofdiastereomers) 90 Ms-D-Ser CH₂CH₂NHSO₂-(5-(4- 17 741 (Bn)formylphenyl)thiophene-2-yl) 91 Ms-D-Ser CH₂CH₂NHSO₂(4-(2- 19 758 (Bn)(MeOCH₂)PyrrolidinylCH₂)Ph) 92 Ms-D-Ser(CH₂)₅NHSO₂(5-pyrid-2-ylthiophene-2- 12 756 (Bn) yl) (Mixture ofdiastereomers) 93 Ms-D-Ser CH₂CH₂NHSO₂(5-(2- 40 812 (Bn)(morpholinoCH₂)phenyl)thiophene-2-yl) 94 Ms-D-Ser CH₂CH₂NHSO₂(5-(4- 22812 (Bn) (morpholinoCH₂)phenyl)thiophene-2-yl) 95 Ms-D-SerCH₂CH₂NHSO₂(5-(3- 30 810 (Bn) (piperidinylCH₂)phenyl)thiophene-2-yl) 96Ms-D-Ser CH₂CH₂NHSO₂(2-acetamido-4- 23 709 (Bn) methylthiazol-5-yl) 97Ms-D-Ser CH₂CH₂NHSO₂(1- 32 671 (Bn) phenylsulfonylpiperidin-4-yl) 98Ms-D-Ser CH₂CH₂NHSO₂-(5-(2- 24 741 (Bn) formylphenyl)thiophene-2-yl) 99Ms-D-Ser CH₂CH₂NHSO₂((CH₃O)CH₃NCH₂Ph) 21 704 (Bn) (Mixture ofdiastereomers) 100 Ms-D-Ser CH₂CH₂NHSO₂(4-ethylpiperazin-1- 21 756 (Bn)yl)CH₂Ph) (Mixture of diastereomers) 101 Ms-D-Ser CH₂CH₂NHSO₂(5-(3- 22798 (Bn) (Et₂NCH₂)phenyl)thiophene-2-yl) 102 Ms-D-Ser CH₂CH₂NHSO₂(5-(3-36 838 (Bn) (Cyclohexyl(Me)NCH₂)phenyl)thiophene- 2-yl) 103 Ms-D-SerCH₂CH₂NHSO₂(5-(3- 24 796 (Bn) (pyrrolidinylCH₂)phenyl)thiophene-2-yl)104 Ms-D-Ser CH₂CH₂NHSO₂(5-(3- 10 738 (Bn) cyanophenyl)thiophene-2-yl)105 Ms-D-Ser CH₂CH₂NHSO₂(4-(4- 14 816 (Bn) acetamidophenoxy)CH₂Ph) (M +Na) 106 Ms-D-Ser CH₂CH₂NHSO₂(5-(3- 44 782 (Bn)(azetidinylCH₂)phenyl)thiophene-2-yl) 107 Ms-D-Ser1-(5-pyridin-2-ylthiophene-2-yl- 23 754 (Bn) SO₂)Piperidin-4-yl)(Mixture of diastereomers) 108 Ms-D-Ser CONHCH₂CH₂NHSO₂(5-(3-(N-ethyl-N-10 784 (Bn) methylaminomethyl)phenyl)thiophene- 2-yl) 109 Ms-D-SerCH₂CH₂NHSO₂(5-(3-(bis(2- 22 858 (Bn)methoxyethyl)aminomethyl)phenyl)thiophene-2- yl) 110 Ms-D-SerCH₂CH₂NHSO₂(5-(3-cyanophenyl)thiophene-2- 11 738 (Bn) yl) (Mixture ofdiastereomers) 111 Ms-D-Ser CH₂CH₂NHSO₂(4-(3-pyrrolin-1-yl)CH₂Ph) 73 712(Bn) (Mixture of diastereomers) 112 Ms-D-SerCH₂CH₂NHSO₂((4-(CH₃SO₂)piperazin-1- 37 807 (Bn) yl)CH₂Ph) 113 Ms-D-SerCH₂CH₂NHSO₂((4-pyrimid-2-yl)piperazin-1- 24 807 (Bn) yl)CH₂Ph 114Ms-D-Ser CH₂CH₂NHSO₂(5-(3- 33 828 (Bn)(thiomorpholinoCH₂)phenyl)thiophene-2-yl) 115 Ms-D-SerCH₂CH₂NHSO₂(5-(3-(4- 16 824 (Bn)ketopiperidinylCH₂)phenyl)thiophene-2-yl) 116 Ms-L-Ser CH₂CH₂NHSO₂Ph 100631 (Bn)

[0147] TABLE 5 Inhibitory Activity of Achiral P₂ Mimetic α-Ketoamides

Ex. Calpain Synthesis MS No. Q R IC₅₀ nM Method (M + 1)* 117 Benzoyl

800 A 563 118 2,6-Dichlorobenzoyl

36 A  631, 633 119 2,6-Dichloro-3- methylbenzoyl

61 A 645 120 2,6-Difluorobenzoyl

20 A 599 121 2,4,6-Trifluorobenzoyl

85 A 618 122 2,3,4,5,6-Pentafluoro- benzoyl

28 A 653 123 3,4-Methylenedioxy- benzoyl

>1000 A 607 124 2,5-Dichlorobenzoyl

68 A  631, 633 125 2-Chloro-5- methoxybenzoyl

65 A 627 126 3,5-bis(trifluoro- methyl)benzoyl

600 A 699 127 2,6-Dimethylbenzoyl

178 A 591 128 2,6-Dichloronicotinoyl

80 A  634, 636 129 2,6-Dichlorobenzoyl

21 A  658, 660 130 2,6-Dichlorobenzoyl

20 A  687, 689 131 2,6-Dichlorobenzoyl

29 A  729, 731 132 2,6-Dichlorobenzoyl

83 A  723, 725 133 2,6-Dichlorobenzoyl

11 A  655, 657 134 2,6-Dichlorobenzoyl

100 A  688, 690 135 2,6-Difluorobenzoyl

22 A 645 (M + Na) 136 2,6-Difluorobenzoyl

62 A 611 (M + Na) 137 2,6-Diethylbenzoyl

145 A 631 (M + Na) 138 2,6-Dimethoxybenzoyl

4000 A 613 139 2-Isopropylbenzoyl

168 A 595 140 2-Chloro-6- fluorobenzoyl

58 A 605 141 2-Fluoro-6-trifluoro- methylbenzoyl

58 A 639 142 2,3,4,5,6-Pentafluoro- benzoyl

32 A 643 143 2-Methylpropanoyl

1500 A 529 144 3-Methylbutanoyl

590 A 543 145 4-Methpentanoyl

29 A 557 146 3- Cyclopentylpropanoyl

1000 A 583 147 E-3-Hexenoyl

1000 A 555 148 4-Phenylpentanoyl

87 A 641 (M + Na) 149 4-Phenylbutanoyl

1500 A 627 (M + Na) 150 4-Methylpentanoyl

15 A 603 (M + Na) 151 3- Cyclopentylpropanoyl

420 A 607

[0148] As those skilled in the art will appreciate, numerous changes andmodifications may be made to the preferred embodiments of the inventionwithout departing from the spirit of the invention. It is intended thatall such variations fall within the scope of the invention.

[0149] It is intended that each of the patents, applications, andprinted publications mentioned in this specification be herebyincorporated by reference in their entirety.

What is claimed is:
 1. A compound having the Formula I:

wherein: Q has the formula G-B—(CHR⁴)_(v) where R⁴ is independently H oralkyl having from 1 to 4 carbons; v is 0, 1, or 2; B is selected fromthe group consisting of C(═O), OC(═O), S(═O)_(m), CH₂, a bond, NR⁵C(═O),S(═O)_(m)-A-C(═O), and C(═O)-A-C(═O), where R⁵ is H or lower alkyl; m is0, 1, or 2; A is lower alkylene or cycloalkylene, optionally substitutedwith one or more halogen atoms, aryl, or heteroaryl groups; M is acarbon atom; G is selected from the group consisting of H, a blockinggroup, lower alkyl, lower alkenyl, aryl having from about 6 to about 14carbons, heterocyclyl having from about 5 to about 14 ring atoms,heterocycloalkyl having from about 5 to about 14 ring atoms, arylalkylhaving from about 7 to about 15 carbons, heteroarylalkyl, andarylheteroalkyl wherein the aryl portion can be unfused or fused withthe heteroalkyl ring, said alkyl, aryl, heterocyclyl, heterocycloalkyl,arylalkyl, heteroarylalkyl, and arylheteroalkyl groups being optionallysubstituted with one or more J groups; J is selected from the groupconsisting of halogen, CN, nitro, lower alkyl, cycloalkyl,heterocycloalkyl, heteroalkyl, halogenated alkyl, aryloxyalkyl,alkylthio, alkylsulfonyl, aryl, heteroaryl, arylalkyl, arylalkyloxy,arylsulfonyl, heteroarylsulfonyl, alkoxycarbonyl, alkoxyalkyl, acyl,alkoxy, hydroxy, carboxy, hydroxyalkyl, amino, alkylamino, andaminoalkyl, said amino group or said amino group of said aminoalkyl oralkylamino group being optionally substituted with an acyl group, analkoxy group, or with 1 to 3 aryl, lower alkyl, cycloalkyl, oralkoxyalkyl groups; and said aryl, heteroaryl, heterocycloalkyl, andheteroalkyl groups being further optionally substituted by a J group;each Aaa is independently an amino acid which optionally contains one ormore blocking groups; n is 0, 1, 2, or 3; R¹ and R² are independentlyselected from the group consisting of H, alkyl having from one to about6 carbons, arylalkyl having from about7 to about 15 carbons, heteroalkylin which the ring contains from about 5 to about 14 ring atoms,heteroarylalkyl in which the heteroaryl ring contains from about 5 toabout 14 ring atoms, alkoxyalkyl, a side chain of a naturally occurringamino acid in the R or S configuration, and (CH₂)_(p)NH-L, said alkyl,arylalkyl, heteroalkyl, heteroarylalkyl, and alkoxyalkyl groups beingoptionally substituted with one or more J groups; p is 0, 1, 2, or 3; Lis selected from the group consisting of alkoxycarbonyl having from 2 toabout 7 carbons, arylalkoxycarbonyl in which the arylalkoxy groupcontains about 7 to about 15 carbons, and S(═O)₂R⁶; R⁶ is selected fromthe group consisting of lower alkyl, and aryl having from about 6 toabout 14 carbons; R³ is selected from the group consisting of H, alkylhaving from one to about 6 carbons, arylalkyl having from about 7 toabout 15 carbons, heteroalkyl in which the ring contains from about 5 toabout 14 ring atoms, heteroarylalkyl in which the heteroaryl ringcontains from about 5 to about 14 ring atoms, alkoxyalkyl, a side chainof a naturally occurring amino acid in the R or S configuration,(CH₂)_(p)NH-L, C(═O)R⁷, S(═O)₂R⁷, a blocking group, and when combinedwith the carbon atom to which Ris attached an alkylene group having from2 to 5 carbons, said alkylene group being optionally substituted with agroup selected from the group consisting of aryl, azide, CN, a protectedamino group, and OSO₂-aryl, said alkyl, arylalkyl, heteroalkyl,heteroarylalkyl, and alkoxyalkyl groups being optionally substitutedwith one or more J groups; R⁷ is selected from the group consisting ofaryl having from about 6 to about 14 carbons, heteroaryl having fromabout 5 to about 14 ring atoms, arylalkyl having from about 7 to about15 carbons, alkyl having from 1 to about 10 carbons, said aryl,heteroaryl, arylalkyl and alkyl groups being optionally substituted withone or more J groups, heteroalkyl having from 2 to about 7 carbons,alkoxy having from about 1 to about 10 carbons, and amino optionallysubstituted with 1 or more alkyl groups; q is 0 or 1; Z is selected fromthe group consisting of C (═O)C(═O)NH—X-A¹-K and

 X is a bond or —O—;  A¹ is the same as A;  K is selected from the groupconsisting of N(R¹⁰) Y,

 and SO₂N(R⁸)(R¹⁰);  D is a fused aryl or heteroaryl group;  R¹¹ isselected from the group consising of alkoxy, aryloxy, and NHR¹²;  R¹² isselected from the group consisting of H, alkyl, aryl, and heteroaryl,said alkyl, aryl or heteroaryl groups being optionally substituted withone or more J groups;  Y is selected from the group consisting of SOR⁸,C(═O)NHR⁹, C(═S)NHR⁹, C(═NCN)R¹¹, C(═NC(═O)NHR¹⁰)R¹¹, and CO₂R⁸;  R⁸ isselected from the group consisting of alkyl, alkoxy, aryl, andheterocyclyl, said alkyl, alkoxy, aryl, or heterocyclyl groups beingoptionally substituted with one or more J groups;  R⁹ is selected fromthe group consisting of H, alkyl, aryl, and heteroaryl, said alkyl,aryl, or heteroaryl groups being optionally substituted with one or moreJ groups;  or an R⁹ alkyl group may be combined with an A¹ alkylenegroup to form a N-containing heterocyclic 5- or 6-membered ring;  R¹⁰ isselected from the group consisting of H and lower alkyl;  or in themoiety SO₂N(R⁸)R¹⁰, R⁸ and R¹⁰ may be combined together with the N atomto which they are attached to form a N-containing heterocyclic 5- or6-membered ring;  or where A¹ is an alkylene group, and K is N(R¹⁰)Ywherein R¹⁰ is alkyl, said R¹⁰ alkyl group may be combined with said A¹alkylene group to form a N-containing heterocyclic 5- or 6-memberedring;  or a pharmaceutically acceptable salt thereof.
 2. The compound ofclaim 1 wherein n and v are each 0, q is 1, B is a bond, and G is H. 3.The compound of claim 1 wherein R¹ is the sidechain of a naturallyoccurring amino acid.
 4. The compound of claim 1 wherein R³ is—S(═O)₂R⁷.
 5. The compound of claim 1 wherein R² is benzyl oralkoxyalkyl.
 6. The compound of claim 1 wherein X is a bond, and Y isSO₂R⁸.
 7. The compound of claim 1 wherein A¹ is —CH₂—CH₂—,—CH₂—CH(CH₃)—, or —(CH₃)CH—CH₂—.
 8. The compound of claim 1 wherein R¹is a serine sidechain, which is optionally capped with a benzyl group.9. The compound of claim 8 wherein M is a carbon atom in the Dconfiguration.
 10. The compound of claim 1 wherein R² is benzyl, R⁷ ismethyl, and R⁸ is substituted phenyl, unsubstituted phenyl, substitutedheteroaryl, or unsubstituted heteroaryl.
 11. The compound of claim 1wherein R⁸ is aryl, aryl substituted with amino, aryl substituted withheterocyclomethyl, heteroaryl, alkyl substituted with heteroaryl, orheteroaryl substituted with alkylthio, haloalkyl, alkyl, phenylsulfonyl,halogen, aminophenyl, amino, or dialkylaminoalkyl.
 12. The compound ofclaim 1 wherein n and v are each 0, q is 1, R¹ is the side chain of anamino acid in the D- or L-configuration, R³ is S(═O)₂R⁷, G is H, B is abond, R² is benzyl or alkoxyalkyl, X is a bond, and Y is SO₂R⁸.
 13. Thecompound of claim 1 wherein A¹ is CH₂CH₂, CH₂CH(CH₃), or (CH₃)CHCH₂. 14.The compound of claim 1 wherein R¹ is a serine side chain in theD-configuration in which the hydroxyl group is capped with benzyl, R²isbenzyl, R⁷ is methyl, and R⁸ is substituted or unsubstituted phenyl orsubstituted or unsubstituted heteroaryl.
 15. The compound of claim 1wherein R₁-R₄, B, G, Aaa, X, A¹, Y, n, q and v are selected inaccordance with Tables 2 and
 3. 16. The compound of claim 1 whereinR₁-R₄, B, G, Aaa, X, A¹, Y, n, q and v are each independently selectedfrom the group of substituents shown in Tables 2 and
 3. 17. The compoundof claim 1 having the Formula:

wherein: W, R₂ and R are independently selected from the group ofsubstituents shown in Table
 2. 18. The compound of claim 17 wherein W,R₂ and R are selected in accordance with Table
 2. 19. The compound ofclaim 1 having the Formula:

wherein D is CH₂CH₂N(CH₃)SO₂Ph or has one of the formulas:


20. The compound of claim 1 having the formula:

wherein W and R are independently selected from the group ofsubstituents shown in Table
 4. 21. The compound of claim 20 wherein Wand R are selected in accordance with Table
 4. 22. The compound of claim1 having the Formula:

wherein Q and R are independently selected from the group ofsubstituents shown in Table
 5. 23. The compound of claim 22 wherein Qand R are selected in accordance with Table
 5. 24. The compound of claim1 wherein n, v and q are each 0; B is (C═O); and G is phenyl or loweralkyl, said phenyl or lower alkyl groups being optionally substitutedwith one or more J groups.
 25. A composition for inhibiting a serineprotease or a cysteine protease comprising a compound of claim 1 and apharmaceutically acceptable carrier.
 26. A method for inhibiting aserine protease or a cysteine protease comprising contacting a proteaseselected from the group consisting of serine proteases and cysteineproteases with an inhibitory amount of a compound of claim 1.